683 MycoKeys
MycoKeys 106: 225-250 (2024)
DOI: 10.3897/mycokeys.106.122890
Research Article
New species and records of Botryosphaeriales
(Dothideomycetes) associated with tree dieback in Beijing, China
Yingying Wu™®, Cheng Peng’, Rong Yuan'®, Mingwei Zhang', Yang Hu2, Chengming Tian'®
1 The Key Laboratory for Silviculture and Conservation of the Ministry of Education, Beijing Forestry University, Beijing 100083, China
2 The Forestry Protection Station of Tonghzou Strict in Beijing, Beijing 101100, China
Corresponding author: Chengming Tian (chengmt@bjfu.edu.cn)
OPEN Qaceess
Academic editor: Ning Jiang
Received: 13 March 2024
Accepted: 2 June 2024
Published: 27 June 2024
Citation: Wu Y, Peng C, Yuan R, Zhang
M, Hu Y, Tian C (2024) New species
and records of Botryosphaeriales
(Dothideomycetes) associated
with tree dieback in Beijing, China.
Mycokeys 106: 225-250. https://doi.
org/10.3897/mycokeys. 106.122890
Copyright: © Yingying Wu et al.
This is an open access article distributed under
terms of the Creative Commons Attribution
License (Attribution 4.0 International - CC BY 4.0).
Abstract
Botryosphaeriales species are important pathogens that have worldwide distribution.
In this study, 23 Botryosphaeriales strains were isolated from 13 host species during
a dieback disease survey in Beijing, China. Based on morphological and phylogenet-
ic analyses, six Botryosphaeriales species were identified, including two new species
named Dothiorella hortiarborum sp. nov. and Phaeobotryon fraxini sp. nov., and four new
host records: Aplosporella ginkgonis from Cotinus coggygria var. cinereus, A. javeedii
from Acer miyabei, Acer truncatum, Forsythia suspensa, Lagerstroemia indica, Sambucus
williamsii, Syringa vulgaris, Ulmus pumila, Xanthoceras sorbifolium, A. yangingensis from
Acer truncatum, and Do. acericola from Forsythia suspensa, Ginkgo biloba, and Syringa
oblata. This study enriches the species diversity associated with tree dieback in Beijing,
China, and contributes to the further study of the taxonomy of this order.
Key words: Dothiorella, morphology, Phaeobotryon, phylogeny, taxonomy
Introduction
Botryosphaeriales species are important plant pathogens commonly found
on the trunks and branches of woody plants (Phillips et al. 2013; Lawrence et
al. 2017; Zhu et al. 2018; Zhang et al. 2021). They are associated with branch
canker, dieback, decline, and death, with consequences for the ecological and
economic value of the forest (Slippers and Wingfield 2007; Phillips et al. 2013;
Mohali-Castillo 2023). Botryosphaeriales species occur on a wide range of
hosts, in the form of endophytes on woody plants and herbs, lichens, and even
seaweed leaves in marine environments, suggesting that they have great po-
tential for research value (Yang et al. 2017; Akinsanmi et al. 2019; Zhang et al.
2021; Mohali-Castillo 2023; Rathnayaka et al. 2023).
Phylogenetic analyses of DNA sequence data have an enormous influence
on the systematics and taxonomy of the order Botryosphaeriales, including re-
defining families and genera and identifying new species (Phillips et al. 2019;
Mohali-Castillo 2023). Schoch et al. (2006) combined SSU, LSU, tef7-a, and
rpb2 to first propose the order Botryosphaeriales, which contains only a sin-
gle family of Botryosphaeriaceae. Minnis et al. (2012) supplemented the DNA
sequence data of Planistromellaceae with phylogenetic analyses combining
225
Yingying Wu et al.: New species and records of Botryosphaeriales
SSU, ITS, LSU, and rpb7, which introduced the family into the Botryosphaeri-
ales. Wikee et al. (2013) reintroduced the Phyllostictaceae, grouped under
Botryosphaeriales, to accommodate Phyllosticta using intronic genes (ITS, act,
and tefl-a) and highly conserved coding regions of genes (LSU and GPDH).
Slippers et al. (2013) added three new families, Aplosporellaceae (Aplosporella
and Bagnisiella), Melanopsaceae (Melanops), and Saccharataceae (Sacchara-
ta), to Botryosphaeriales based on DNA sequence data of six loci (SSU, LSU,
ITS, tef1-a, tub2, and mtSSU). Wyka and Broders (2016) introduced Septori-
oideaceae based on morphological and molecular evidence. Yang et al. (2017)
mentioned that the LSU-rpb2 combination could effectively classify taxa at
the family and genus levels, and rpb2 in combination with ITS, tef7-a, and tub2
added additional resolution for species delimitation. For this reason, they com-
bined the five fragments ITS, tef1-a, tub2, LSU, and rpb2 to propose two new
families, Endomelanconiopsisaceae and Pseudofusicoccumaceae. Therefore,
Botryosphaeriales contained a total of nine families. However, Phillips et al.
(2019) reassessed the families of Botryosphaeriales in terms of morphology
of the sexual morphs and phylogenetic relationships of ITS and LSU sequence
data, ultimately concluding that the order contained only six families (Aplospo-
rellaceae, Botryosphaeriaceae, Melanopsaceae, Phyllostictaceae, Planistro-
mellaceae, and Saccharataceae), with Endomelanconiopsisaceae, Pseudofu-
sicoccumaceae, and Septorioideaceae as synonyms of existing families. Up
to date, six families and 32 genera are accepted in Botryosphaeriales (https://
www.outlineoffungi.org/). Of these, Botryosphaeriaceae is rich in species diver-
sity, high in pathogenicity, and widely distributed.
Botryosphaeriaceae was first established by Theissen and Sydow (1918),
containing three genera: Botryosphaeria, Dibotryon, and Phaeobotryon. Mor-
phologically, Botryosphaericeae species are distinctive from other families
in Botryosphaeriales by their large, ovoid to oblong, usually hyaline, aseptate
ascospores (Phillips et al. 2013). Liu et al. (2012) assumed that ascospores
could become pigmented and septate with age. Conidia in the asexual state of
Botryosphaericeae are diverse in morphological characteristics (Phillips et al.
2005). Phylogenetically, however, there is a random distribution of hyaline or
colored conidia or ascospores in the phylogenetic tree of Botryosphaericeae
(Slippers et al. 2013). Therefore, accurate identification of species in the family
by a single circumscription is not suitable. Currently, 22 genera and more than
200 species are contained within the family (https://www.outlineoffungi.org/).
Recently, many new species have been introduced in the Botryosphaeriaceae,
especially in the genera Dothiorella and Phaeobotryon (Jia et al. 2023; Li et al.
2023; Lin et al. 2023a; Wu et al. 2023).
Saccardo (1880) first established Dothiorella and designated Do. pyrenophora
as the type species. Up to now, some scholars have made systematic revisions
of Dothiorella to establish a more stable phylogenetic relationship (Dissanayake
et al. 2016; Dissanayake et al. 2020; Zhang et al. 2021). The distinctive features
of the genera are that the conidia are colored in the early stages of development,
and with 1-septate, the sexual form of ascospores is brown and septate (Sena-
nayake et al. 2023). The type species of the genus Phaeobotryon is P. cercidis,
which is characterized by 2-septate brown ascospores with conical apiculate-el-
liptic to oblong or obovoid shapes at both ends and hyaline or brown conidia
(Phillips et al. 2013; Fan et al. 2015b; Zhu et al. 2018; Pan et al. 2019).
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890 296
Yingying Wu et al.: New species and records of Botryosphaeriales
In recent years, multiple studies have revealed that new species of Botryos-
phaeriales infest branches and trunks. Pan et al. (2019) found that Phaeobo-
tryon rhois and Diplodia quercicola were detrimental to Rhus typhina and Quer-
cus variabilis separately in Yudu Mountain, Beijing. Aplosporella yangingensis
and Dothiorella baihuashan are mainly recorded on Pinaceae or Cupressaceae
(Lin et al. 2023a). Lasiodiplodia regiae caused the canker and dieback of ap-
ple trees (Wang et al. 2023). These studies suggest that Botryosphaeriales is
rich in species diversity and has the potential to continue to be explored for
new species. During the investigation of plant pathogens in Beijing, a higher
number of diseased plant branches caused by Botryosphaeriales fungi were
found. This study used phylogenetic analysis and morphological comparisons
to describe new species and new host records, enriching the fungal taxa with-
in Botryosphaeriales.
Materials and method
Sample collection and fungal isolation
A survey on dieback diseases was conducted from March to November 2023
in the Tongzhou District of Beijing, China. A total of thirteen tree species were
examined, namely Acer miyabei, A. truncatum, Cotinus coggygria var. cinere-
us, Forsythia suspensa, Fraxinus chinensis, Ginkgo biloba, Lagerstroemia indica,
Sambucus williamsii, Styphnolobium japonicum, Syringa oblata, Syringa vulgaris,
Ulmus pumila, and Xanthoceras sorbifolium. Twenty specimens showing typical
dieback symptoms (Fig. 1) with typical conidiomata and/or ascomata were col-
lected. All samples were placed in paper bags and transported to the laboratory.
Specimens with typical conidiomata pycnidial were selected for isolation. Re-
moving the spore mass from conidiomata and generating single spore colonies
or plating superficially sterilized diseased tissue on potato dextrose agar plates
(PDA; containing 200 g potatoes, 20 g dextrose, and 20 g agar per liter) and in-
cubating Petri dishes at 25 °C in the dark for 2-3 d. When colonies just formed,
they transferred to fresh PDA Petri dishes (Crous et al. 2019). All specimens
were deposited at the Museum of Beijing Forestry University (BUFC), and all
cultures were preserved at the China Forestry Culture Collection Center (CFCC).
Morphological observation
Cultures were incubated on PDA at 25 °C in a 12-h day/night regime (Crous et
al. 2019). After 14 days, the colonies were measured, and characteristics based
on the color, shape, and sparseness of the aerial mycelium of the pathogen
colonies were observed and recorded. Conidiomata were manually sectioned
with a double-edged razor blade. Observations were conducted using a Leica
DM 2,500 dissecting microscope (Wetzlar, Germany) and a Nikon Eclipse 80i
compound microscope, equipped with differential interference contrast (DIC)
illumination. Images were captured using a Nis DS-Ri2 camera with the Nikon
Nis-Elements F4.30.01 software. Conidial length was measured from the base
of the basal cell to the base of the apical appendage, while conidial width was
measured at its widest point. A randomized selection of conidia was used for
measurement (n = 50).
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890 907
Yingying Wu et al.: New species and records of Botryosphaeriales
Figure 1. Disease symptoms associated with Botryosphaeriales species collected from Tongzhou District, Beijing, China
A Xanthoceras sorbifolium B Fraxinus chinensis C Lagerstroemia indica D Sambucus williamsii E Styphnolobium japoni-
cum F Forsythia suspensa.
DNA extraction, PCR amplification, and sequencing
Genetic DNA was extracted using the cetyltrime-thylammonium bromide
(CTAB) method when the mycelium was well spread on the PDA. DNA sam-
ples were stored at -20 °C. The PCR reaction primers (forward and reverse)
and amplification conditions are detailed in Table 1. Polymerase chain reaction
(PCR) amplification was run on a PTC-200 Thermal Cycler amplifier from Bio-
Rad, USA. The PCR amplification systems were all 20 uL, including 10 uL of Mix
(Promega), 7 UL of double deionized water, 1 pL each of pre- and post-primers,
and 1 uL of DNA template. PCR products were assayed by electrophoresis on
2% agarose gels. Amplified PCR products were sent to a commercial sequenc-
ing provider (Tsingke Biotechnology Co. Ltd., Beijing, China).
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890 228
Yingying Wu et al.: New species and records of Botryosphaeriales
Table 1. Genes used in this study with PCR primers.
Locus PCR primers PCR: thermal cycles: (Annealing temp. in bold) References
ITS ITS1/ITS4 (95 °C: 30s, 51 °C: 30 s, 72 °C: 1 min) x 35 cycles White et al. 1990
LSU LROR/LR5 (95 °C: 45s, 55 °C: 30s, 72°C: 1 min) x 35 cycles | Vilgalys and Hester 1990
tefl-a | EF1-728F/EF1-986R | (95 °C: 15s, 55 °C: 30s, 72 °C: 1 min) x 35 cycles | Carbone and Kohn 1999
tub2 Bt2a/Bt2b (95 °C: 30 s, 55 °C: 30 s, 72 °C: 1 min) x 35 cycles | Glass and Donaldson 1995
Phylogenetic analyses
The sequences obtained were assembled using SeqMan v. 7.1.0 software,
and reference sequences from related publications (Phillips et al. 2019; Li
et al. 2023; Lin et al. 2023a; Wu et al. 2023) were retrieved from the National
Center for Biotechnology Information (NCBI; https://www.ncbi.nIm.nih.gov).
All sequences generated in this study were submitted to GenBank (Table 2).
Sequences were aligned in MAFFT v. 7 at the web server (https://mafft.cbre.
jp/alignment/server/) (Katoh and Standley 2013; Katoh et al. 2019) and further
adjustments and editing of the sequences were made with MEGA v. 6 (Tamura
et al. 2013). Maximum parsimony (MP), maximum likelihood (ML), and Bayes-
ian inference (BI) were selected to construct phylogenetic trees using PAUP
v. 4.0610, PhyML 3.0, and MrBayes V3.1.2 (Huelsenbeck and Ronquist 2001;
Swofford 2003; Silvestro and Michalak 2012). Phylograms were visualized with
FigTree v. 1.4.0 (http://tree.bio.ed.ac.uk/software/figtree/) and additional ed-
ited with Adobe Illustrator CS v. 5 (Adobe Systems Inc., USA). Maximum-par-
simony bootstrap values (MPBP) and maximum-likelihood bootstrap values
(MLBP) = 50% and Bayesian posterior probabilities (BYPP) = 0.90 are shown
for each tree.
Maximum parsimony analysis was performed using the tree bisection and
reconnection (TBR) branch swapping algorithm with a heuristic search op-
tion of 1000 random-addition sequences (Swofford 2003). Max trees were
set to 5000 branches of zero length, and all parsimonious trees were saved.
Other measures calculated were tree length (TL), consistency index (Cl),
retention index (RI), and rescaled consistency (RC) (Swofford 2003). Maxi-
mum likelihood analysis was performed with the GTR GAMMA model of site
substitution, including estimation of gamma-distributed rate heterogeneity
and a proportion of invariant sites (Guindon et al. 2010). The branch support
from MP and ML analysis was evaluated with a bootstrapping (BS) method
of 1 000 replicates (Hillis and Bull 1993). The Bayesian inference analysis
employing a Markov chain Monte Carlo (MCMC) algorithm was performed
with Bayesian posterior probabilities (Rannala and Yang 1996). The mod-
el of nucleotide substitution was estimated by MrModeltest v.2.3 (Posada
and Crandall 1998), and a weighted Bayesian analysis was considered. Two
MCMC chains were run starting from random trees for 1,000,000 genera-
tions and stopped when the average standard deviation of split frequencies
fell below 0.01; the trees were sampled every 100" generation. The first
25% of trees were discarded as the burn-in phase of each analysis, and the
Bayesian posterior probabilities (BPP) were calculated using the remaining
7,500 trees.
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890 999
Yingying Wu et al.: New species and records of Botryosphaeriales
Table 2. Isolates of Aplosporella, Dothiorella, and Phaeobotryon used in the molecular analyses in this study. Notes: NA:
not applicable, Strains in this study are marked in bold, T: ex-type strains.
Species
Aplosporella africana
A. africana
A. artocarpi
A. ginkgonis
A. ginkgonis
A. ginkgonis
A. hesperidica
A. hesperidica
A. javeedii
A. javeedii
A. javeedii
A. javeedii
A. javeedii
A. javeedii
A. javeedii
A. javeedii
A. javeedii
A. javeedii
A. javeedii
A. javeedii
A. javeedii
A. javeedii
A. javeedii
A. javeedii
A. macropycnidia
A. macropycnidia
A. papillata
A. papillata
A. prunicola
A. prunicola
A. sophorae
A. thailandica
A. yalgorensis
A. yalgorensis
A. yangingensis
A. yangingensis
A. yangingensis
A. yanqgingensis
Alanomyces indica
Dothiorella alpina
Do. acacicola
Do. acericola
Do. acericola
Do. acericola
Do. acericola
Strain
CBS 121777°
CBS 1217778"
CPC 22791"
CFCC 52442"
CFCC 89661"
CFCC 70746
CBS 732.79"
CBS 208.37
CFCC 50054"
CFCC 50052
CFCC 58330
CFCC 58329
CFCC 58412
CFCC 70733
CFCC 70734
CFCC 70735
CFCC 70736
CFCC 70737
CFCC 70739
CFCC 70740
CFCC 70741
CFCC 70742
CFCC 70744
CFCC 70745
CGMCC 3.17725"
CGMCC 3.17726
CBS 121780°
CBS 121781
CBS 121167°
STE-U 6326
CPC 29688"
MFLU 16-0615"
MUCC5117
MUCC512
CFCC 587917
CFCC 58792"
CFCC 70743
CFCC 70738
CBS 134264"
CGMCC 3-180017
CBS 141295'
KUMCC 18-0137"
CFCC 70755
CFCC 70760
CFCC 70761
Host
Acacia mellifera
Acacia mellifera
Artocarpus
heterophyllus
Rhus typhina
Rhus typhina
Cotinus coggygria var.
cinereus
Citrus aurantium
Citrus sinensis
Juniperus chinensis
Gleditsia sinensis
Populus canadensis
Populus beijingensis
Populus alba var.
pyramidalis
Styphnolobium
japonicum
Forsythia suspensa
Forsythia suspensa
Ulmus pumila
Acer truncatum
Sambucus williamsii
Acer miyabei
Lagerstroemia indica
Xanthoceras
sorbifolium
Syringa vulgaris
Ulmus pumila
Cerasus yedoensis
Cerasus yedoensis
Acacia tortillas
Acacia tortillas
Prunus persica var.
nucipersica
Prunus persica var.
nucipersica
Sophora microphylla
Dead stems
Acacia cochlearis
Eucalyptus
gomphocephala
Platycladus orientalis
Platycladus orientalis
Acer truncatum
Acer truncatum
Soil
Platycladus orientalis
Acacia mearnsii
Acer palmatum
Forsythia suspensa
Ginkgo biloba
Syringa oblata
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890
Origin
Namibia
Namibia
Thailand
China
China
China
Buenos Aires
Zimbabwe
China
China
China
China
China
China
China
China
China
China
China
China
China
China
China
China
China
China
South Africa
South Africa
South Africa
South Africa
New Zealand
North
Thailand
Australia
Australia
China
China
China
China
India
China
Réunion
China
China
China
China
ITS
KF766196
EU101316
KM006450
MH133916
KM030583
PP188498
KX464083
JX681069
KP208840
KP208838
0Q651161
0Q651162
0Q651163
PP188499
PP188500
PP188501
PP188502
PP188503
PP188504
PP188505
PP188506
PP188507
PP188508
PP188509
KT343648
KT343649
EU101328
EU101329
KF766147
EF564375
KY173388
KX423536
EF591926
EF591927
0Q651164
0Q651165
PP188510
PP188511
HF563622
KX499645
KX228269
MK359449
PP188520
PP188521
PP188522
GenBank accession numbers
tef1-a
EU101360
EU101361
KM006481
MH133950
KM030597
PP541796
NA
NA
KP208846
KP208844
0Q692921
0Q692922
0Q692923
PP541797
PP541798
PP541799
PP541800
PP541801
PP541802
PP541803
PP541804
PP541805
PP541806
PP541807
KX011176
KX011177
EU101373
EU101374
NA
NA
NA
KX423537
EF591977
EF591978
0Q692924
0Q692925
PP541808
PP541809
AB872219
KX499651
KX228376
MK361182
PP766251
PP766252
PP766253
tub2
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
PP566659
PP566660
PP566661
LSU
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
230
Yingying Wu et al.: New species and records of Botryosphaeriales
Species Strain Host
Do. albiziae MFLUCC 22-0057' Albizia lebbeck
Do. alpina CFCC 58299' Populus szechuanica
Do. americana CBS 128309' Vitis species and Vitis
vinifera
Do. baihuashanensis CFCC 58549" Juniperus chinensis
Do. baihuashanensis CFCC 58788" Juniperus chinensis
Do. brevicollis CBS 130411 = CMW 36463' Acacia karroo
Do. californica CBS 119635 Laurus nobilis
Do. californica CBS 141587 Umbellularia californica
Do. camelliae CMGCC 3.24158" Camellia oleifera
Do. capri-amissi CBS 121763 = CMW 25403 Acacia erioloba
= CAMS 11587
Do. capri-amissi CBS 121878 = CMW 25404 Acacia erioloba
= CAMS 11597
Do. casuarinae CBS 120688 = CMW 4855" Casuarina sp.
Do. casuarinae CBS 120689 = CMW 4856 Casuarina sp.
Do. casuarinae CBS 120690 = CMW 4857 Casuarina sp.
Do. citricola CBS 124728 = ICMP 16827 Citrus sinensis
Do. citricola CBS 124729 = ICMP 16828" Citrus sinensis
Do. citrimurotticola BES = CGMCC3.20392' Citrus unshiu
Do. citrimurotticola BE8 = CGMCC3.20394 Citrus reticulatachen x
C. sinensis
Do. diospyricola CBS 145972 Diospyros
mespiliformis
Do. dulcispinae CBS 121764 = CMW 25406 Acacia mellifera
= CAMS 1159
Do. dulcispinae CBS 130413 = CMW 36460° Acacia karroo
Do. eriobotryae CBS 140852° Eriobotrya japonica
Do. franceschinii CBS 147722 Rhamnus alaternus
Do. guttulata MFLUCC 17-0242 Alnus glutinosa
Do. heterophyllae CMW 46458° Acacia heterophylla
Do. hortiarborum CFCC 70756" Fraxinus chinensis
Do. hortiarborum CFCC 70757 Fraxinus chinensis
Do. hortiarborum CFCC 70758 Lagerstroemia indica
Do. hortiarborum CFCC 70759 Lagerstroemia indica
Do. iberica CBS 113188 = DA-1 Quercus suber
Do. iberica CBS 113189 = DE-14 Quercus ilex
Do. iberica CBS 115041 = CAP 145° Quercus ilex
Do. irannica CBS 124722 = CJA 153 = Olea europaea
IRAN 1587C™
Do. koae CMW 48017' Acacia koa
Do. lampangensis MFLUCC 18-0232' Rutaceae
Do. longicollis CBS 122066 = CMW 26164 Terminalia sp.
Do. longicollis CBS 122067 = CMW 26165 Lysiphyllum
cunninghamii
Do. longicollis CBS 122068 = CMW 26166' Lysiphyllum
cunninghamii
Do. magnoliae CFCC51563" Magnolia grandiflora
Do. mangifericola CBS 124727' Mangifera indica
Do. mangifericola IRAN 1584C Mangifera indica
Do. moneti WAC 13154 = MUCC 505° Acacia rostellifera
Do. neclivorem DAR 80992" Vitis vinifera
Do. oblonga CBS 121765 = CMW 25407 Acacia mellifera
= CAMS 11627
Do. oblonga CBS 121766 = CMW 25408 Acacia mellifera
= CAMS 1163
Do. obovata MFLUCC22-0058" Pavonia odorata
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890
Origin
Thailand
China
USA: Missouri
China
China
South Africa
Turkey
USA
China
South Africa
South Africa
Australia
Australia
Australia
New Zealand
New Zealand
China
China
South Africa
Namibia
South Africa
Spain
Italy
Italy
Réunion
China
China
China
China
Spain
Spain
Spain
Iran, Golestan
Hawaiian Is.
Thailand
Australia
Australia
Australia
China
Iran
Iran
Australia
Australia
South Africa
South Africa
Thailand
ITS
ON751762
0Q651166
HQ288218
0Q651167
0Q651168
JQ239403
MT587396
KX357188
0Q190531
EU101323
EU101324
DQ846773
DQ846772
DQ846774
EU673322
EU673323
MW880663
MW880661
MT587398
EU101299
JQ239400
KT240287
OP999677
KY797637
MN103794
PP188523
PP188524
PP188525
PP188526
AY573198
AY573199
AY573202
KC898231
MH447652
MK347758
EU144052
EU144053
EU144054
KY111247
KC898221
MT587407
EF591920
KJ573643
EU101300
EU101301
ON751763
tef1-a
ON799588
0Q692932
HQ288262
0Q692933
0Q692934
JQ239390
MT592108
KX357211
0Q241464
EU101368
EU101369
DQ875331
DQ875332
DQ875333
EU673289
EU673290
MW884166
MW884164
MT592110
EU101344
JQ239387
KT240262
0Q067247
NA
MH548348
PP723042
PP723043
PP723044
PP723045
EU673278
AY573230
AY573222
KC898214
MH548338
MK340869
EU144067
EU144068
EU144069
KY213686
KX464614
MT592119
EF591971
KJ573640
EU101345
EU101346
ON799589
GenBank accession numbers
tub2
ON799590
0Q692926
HQ288297
0Q692927
0Q692928
JQ239371
MT592579
KX357165
0Q275064
KX464850
KX464851
DQ875340
DQ875339
DQ875341
KX464852
KX464853
MW884195
MW884193
MT592581
KX464854
JQ239373
MT592582
NA
NA
MH548324
PP566662
PP566663
PP566664
PP566665
EU673097
KX464855
EU673096
KX464856
MH548327
MK412874
KX464857
KX464858
KF 766130
NA
NA
NA
EF591954
KJ577551
KX464862
KX464863
ON799591
LSU
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
NA
231
Yingying Wu et al.: New species and records of Botryosphaeriales
Do.
Do.
Do.
Do.
Do.
Do.
Species
omnivora
omnivora
omnivora
omnivora
omnivora
parva
Do. parva
Do. parva
Do. plurivora
Do. pretoriensis
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do
prunicola
rhamni
rosulata
rosulata
rosulata
rosulata
santali
saprophytica
sarmentorum
sempervirentis
sempervirentis
sp.
sp.
sp.
striata
striata
styphnolobii
symphoricarpicola
tectonae
thailandica
thripsita
ulmacea
uruguayensis
vidmadera
vidmadera
vinea-gemmae
viticola
westralis
. yunnana
Do. yunnana
Do. zanthoxyli
CBS 124721 =
CBS 124730 =
CBS 124731 =
CBS 392.80
CBS 124716 = CJA 241 =
IRAN 1573C
CBS 242.51
CBS 188.87
CBS 124720 = CJA 27 =
IRAN 1579C™
CBS 125580
CBS 124724 = CJA 254 =
IRAN 1557C™
CBS 130404 = CMW 36480°
CBS 124723 = CAP 187 =
IRAN 1541C'
MFLUCC 14-0902"
CBS 121760 = CMW 25389
= CAMS 14447
CBS 121761 = CMW 25392
= CAMS 1147
CBS 121762 = CMW 25395
= CAMS 1150
CBS 500.72
WAC 13155 = MUCC 5097
MFLUCC 23-0210
IMI 63581b
IRAN 1581C = CBS 124719
IRAN 1583C = CBS 124718
= CJA 264°
CBS 121783 = CMW 25432
= CAMS 1187
CBS 121784 = CMW 25430
= CAMS 1185
CBS 121785 = CMW 25433
= CAMS 1188
Cr01"
CPC 33923"
MFLUCC18-0232'
CBS 133991 = CPC 21557 =
MFLUCC 11-0438
CBS 125445 = BRIP 51876a"
CBS 141414"
CBS 124908 = CMW 26763'
CBS 621.74
CBS 725.79"
DAR 81012"
CBS 117009°
WA10NO017
CGMCC 3-17999"
CGMCC 3-18000
CMGCC 3.241597
CJA 35
ICMP 16819
ICMP 16824"
Juglans regia
Juglans regia
Corylus sp.
Corylus sp.
Corylus avellana
Citrus sp.
Acacia karroo
Prunus dulcis
Rhamnus cathartica
Acacia karroo
Acacia mellifera
Acacia mellifera
Medicago sativa
Santalum acuminatum
Ulmus sp.
Cupressus
sempervirens
Cupressus
sempervirens
Acacia mearnsii
Acacia mearnsii
Acacia mearnsii
Citrus sinensis
Citrus sinensis
Styphnolobium
japonicum
Symphoricarpos
Tectona grandis
Dead bamboo culm
Acacia harpophylla
Ulmus laevis
Hexachlamis edulis
Pyrus communis
Pyrus malus
Vitis vinifera
Vitis vinifera
Vitis vinifera
Camellia sp.
Camellia sp.
Zanthoxylum
bungeanum
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890
S
Origin
Iran
Italy
France
Iran
Iran
Austria
Iran
South Africa
Portugal
outh European
Russia
Namibia
South Africa
South Africa
South Africa
Australia
Thailand
UK: England
Iran
Iran
South Africa
South Africa
South Africa
New Zealand
New Zealand
Crym
Italy
Thailand
Thailand
Australia
Germany
Uruguay
Switzerland
Switzerland
Australia
Spain
Australia
China
China
Sichuan
GenBank accession numbers
tef1-a
CBS 124717 = CJA 214 = Juglans regia Iran KC898233 | KC898216 | KX464865
IRAN 1570C
tub2
KC898232 | KC898215 | KX464864
EU673317
EU673316
KC898234
KX464123
KC898217
EU673119
KX464866
KX464867
KC898225 | KC898208 | KX464874
JQ239405 | JQ239392 | JQ239376
EU673313
MF398893
KF766227
EU101293
EU101319
EU673318
EF591924
KX464124 | KX464616 | KX464868
EU673280
MF398945
EU101335
EU101338
EU101364
EU673100
NA
KX464877
KX464878
KX464879
EU673118
EFS91.958
ORS27239 | OR532455 | OR532454
AY573212
EU673102
KC898237 | KC898220 | KX464885
KC898236
EU101333
EU101331
EU101334
EU673320
KC898219
EU101378
EU101376
EU101379
KX464884
KX464859
KX464860
KX464861
NA
MH880849
MK069594
JX646796 | JX646861 | JX646844
MT587415
EU080923
KX464129
KX464130
KJ573644
AY905554
HM009376
KX499643
KX499644
0Q190536
KJ573641
10573681
‘aeons
HM8s00511
0Q241468
KX464888
KJ577552
EU673104
NA
NA
NA
0Q275069
NA
232
Yingying Wu et al.: New species and records of Botryosphaeriales
Species
Neofusicoccum
luteum
Neofusicoccum
parvum
Phaeobotryon
aplosporum
P. aplosporum
P. aplosporum
P. aplosporum
P. aplosporum
P cupressi
P cupressi
P. fraxini
P. fraxini
P juniperi
P juniperi
P mali
P mali
P mali
P mali
P mali
P mamane
P. mamane
P. negundinis
P. negundinis
P. negundinis
P. negundinis
P. negundinis
P. negundinis
P. negundinis
P platycladi
P platycladi
P rhoinum
P rhoinum
P rhois
| P rhois
P rhois
P spiraeae
P spiraeae
P spiraeae
P ulmi
P ulmi
P ulmi
P ulmi
P ulmi
P ulmi
P ulmi
Alanphillipsia
aloeicola
CBS 562.927
CMW 9081'
CFCC 53774
CFCC 53775"
CFCC 53776
CFCC 58596
CFCC 58784
CBS 124700 =
CBS 124701 =
IRAN 1455C™
IRAN 1458C
CFCC 70762"
CFCC 70763
JU001 7
JU005
XJAU 2930"
XJAU 2772
XJAU 2782
XJAU 3094
XJAU 3100
CBS 122980 = CPC 12440"
CPC 12442
CAA 797
CAA 798
CAA 799
CPC 33384
CPC 33388
CPC 34752
MFLUCC 15-0436"
CFCC 58799"
CFCC 58800
CFCC 52449
CFCC 52450°
CFCC 89662 = CCTCC
AF2014017°
94-
1
CBS 114123 = UPSC 2552
CBS 138854 = CPC 24264"
CBS 123.30 = ATCC 24443
CBS 174.63
CMH 299
PB_11f
CBS 138896 = CPC 23674"
Actinidia deliciosa
Populus nigra
Syzygium aromaticum
Rhus typhina
Rhus typhina
Juglans mandshurica
Juglans mandshurica
Cupressus
sempervirens
Cupressus
sempervirens
Fraxinus chinensis
Fraxinus chinensis
Juniperus formosana
Juniperus formosana
Malus pumila
Juglans regia
Malus ‘Royalty’
Elaeagnus angustifolia
Rhus typhina
Sophora chrysophylla
Sophora chrysophylla
Acer negundo
Ligustrum vulgare
Forsythia intermedia
Acer nugundo
Dead stem
Acer negundo
Acer negundo
Platycladus orientalis
Platycladus orientalis
Rhus typhina
Rhus typhina
Rhus typhina
Ulmus pumila
Ulmus laevis
Ulmus sp.
Ulmus glabra
House dust
Ulmus glabra
Aloe sp.
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890
Origin
New Zealand
New Zealand
China
China
China
China
China
Iran
Iran
China
China
China
China
China
China
China
China
China
USA
USA
Russia
Ukraine
Ukraine
Ukraine
Russia
China
China
China
China
ITS
MH862376
AY236943
MN215837
MN215838
0Q651169
0Q651170
FJ919672
FJ919671
PP188527
PP188528
EU673332
EU673333
KX061513
KX464690
KX464968
GenBank accession numbers
LSU
NA
AY236888 | AY236917 NA
MN215836 | MN205996
FJ919661
FJ919660
OP941637 | OP948218
OP941638 | OP948219
0948218 |
MW326854
MW509520 |
MW509518 |
MW326853 | MW509520
0P 948219 |
MW326852
“ws09517
MW326858 | MW509517
MW326878 | MW509518
KX061514 | KX061508
KX061515 | KX061509
MT587542
MT587543 | MT592277
MT587544
KU820970
0Q651172
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
A
N
NA
KM030584 | KM030598
CFCC 89663 = CCTCC Rhus typhina China KM030585 | KM030599 NA
AF2014016
CFCC 586797 Populus alba var. China 0Q651171 | 0Q692929 NA 00652542
pyramidalis
NA
Ulmus glabra MT587539 | MT592273
KP004444 | MT592027
Poland
South Africa
MN215871
MN215872
MN215873
0Q652540
0Q652541
KX464538
KX464539
PP177348
PP177349
0P941644
0P941645
MW367101
MW367094
MW367092
MW367100
MW367093
EU673248
DQ377899
NA
NA
NA
MT587323
MT587324
MT587325
NA
0Q652543
0Q652544
MH133940
MH133941
KM030591
KM030592
NA
MT587320
MT587321
DQ377861
MT587322
NA
NA
KP004472
233
Yingying Wu et al.: New species and records of Botryosphaeriales
Result
Phylogenetic analysis
The BLAST results indicated that the 23 isolates resided in Ap/osporella, Do-
thiorella, and Phaeobotryon (14 for Aplosporella, 7 for Dothiorella, and 2 for
Phaeobotryon). Separate phylogenetic trees for each of the three genera were
constructed in this study.
In Aplosporella, the combined ITS and tef1-a dataset consists of 944 charac-
ters, including alignment gaps (508 for ITS and 436 for tef1-a), of which 794 are
constant and 60 are variable parsimony uninformative characters. MP analysis
with the remaining 90 parsimony-informative characters resulted in one equal-
ly parsimonious tree: tree length (TL) = 230; consistency index (Cl) = 0.817;
retention index (RI) = 0.896; and rescaled consistency index (RC) = 0.732. In
ML analysis based on the combined gene dataset, the matrix had 193 distinct
alignment patterns. Estimated base frequencies are as follows: A = 0.217607,
C = 0.264598, G = 0.259539, T = 0.258256, AC = 2.784746, AG = 2.845183, AT
= 1.353935, CG = 1.848853, CT = 4.935430, GT = 1.000000, gamma distribu-
tion shape parameter: a = 0.157110, and likelihood value of In: -2 499.855852.
The maximum likelihood (ML) and Bayesian methods (BI) for phylogenetic
analyses have the same topology and terminal clades. Fourteen isolates were
distributed in Aplosporella, aggregated with three known species, A. javeedii,
A. yangingensis, and A. ginkgonis, separately (Fig. 2). The single gene tree for
ITS and tef1-a of Aplosporella is shown in Suppl. material 1.
In Dothiorella, sequences of the combined ITS, tef1-a, and tub2 were aligned; the
dataset consists of 1,319 characters, including alignment gaps (534 for ITS, 369
for tef1-a, and 416 for tub2), of which 905 are constant and 107 are variable par-
simony uninformative characters. MP analysis with the remaining 307 parsimo-
ny-informative characters resulted in one equally parsimonious tree: tree length
(TL) = 1,282; consistency index (Cl) = 0.477; retention index (RI) = 0.824; and
rescaled consistency index (RC) = 0.394. In ML analysis based on the combined
gene dataset, the matrix had 601 distinct alignment patterns. Estimated base fre-
quencies are as follows: A = 0.206208, C = 0.312741, G = 0.250328, T = 0.230723,
AC = 0.833804, AG = 2.174710, AT = 1.041501, CG = 0.791470, CT = 3.735830,
GT = 1.000000, gamma distribution shape parameter: a = 0.215045, and likeli-
hood value of In: -8 567.497788. Three of the seven isolates were of the known
species Dothiorella acericola, and the other four isolates formed a separate clade
for designation as new species based on phylogenetic analysis (Fig. 3). The sin-
gle gene tree for ITS, tef7-a, and tub2 of Dothiorella is shown in Suppl. material 2.
In Phaeobotryon, the combined ITS, LSU, and tef7-a dataset consists of 1,394
characters, including alignment gaps (494 for ITS, 333 for LSU, and 567 for
tef1-a), of which 1,218 are constant and 56 are variable parsimony uninforma-
tive characters. MP analysis with the remaining 120 parsimony-informative
characters resulted in one equally parsimonious tree: tree length (TL) = 259;
consistency index (Cl) = 0.799; retention index (RI) = 0.913; and rescaled con-
sistency index (RC) = 0.730. In ML analysis based on the combined gene data-
set, the matrix had 239 distinct alignment patterns. Estimated base frequen-
cies are as follows: A = 0.224820, C = 0.266099, G = 0.277247, T = 0.231833, AC
= 0.602998, AG = 2.181745, AT = 0.500445, CG = 0.607508, CT = 4.549533, GT
= 1.000000, gamma distribution shape parameter: a = 0.020014, and likelihood
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890 034
Yingying Wu et al.: New species and records of Botryosphaeriales
value of In: -3 357.887099. Eight isolates were assigned to Phaeobotryon, one
isolate aggregated with P mali, and two isolates stood alone, not branching off
from known species, representing a new species (Fig. 4). The single gene tree
for ITS, LSU, and tef7-a of Phaeobotryon is shown in Suppl. material 3.
Aplosporella javeedii CFCC 50052
Aplosporella javeedii CFCC 70734
Aplosporella javeediti CFCC 50054
Aplosporella javeedii CFCC 58330
Aplosporella javeedii CFCC 58412
Aplosporella javeedii CFCC 58329
Aplosporella javeedii CFCC 70742
Aplosporella javeedii CFCC 70739
Aplosporella javeedii CFCC 70736
o7/o7/ip APlosporella javeedit CFCC 70744
Aplosporella javeedii CFCC 70741
Aplosporella javeedii CFCC 70737
Aplosporella javeedii CFCC 70735
! Aplosporella javeedii CFCC 70745
Aplosporella javeedii CFCC 70740
Aplosporella javeedii CFCC 70733
Aplosporella sophorae CPC 29688
aia Aplosporella macropycnidia CGMCC 3.17725
| 82/90/0.98] & Aplosporella macropycnidia CGMCC 3.17726
8B/90/I—_ |. Aplosporella africana CBS 121777
74/73/0.98 Aplosporella africana CBS 121778
98/100/1 Aplosporella yalgorensis MUCC 511
Aplosporella yalgorensis MUCC 512
Aplosporella prunicola CBS 121167
?1196/0.99! Anlosporella prunicola STE-U 6326
71/76/0.91 100/100/1; Aplosporella papillata CBS 121780
Aplosporella papillata CBS 121781
Aplosporella yangingensis CFCC 58792
Aplosporella yangingensis CFCC 58791
90/95/- 00/100) 4 nlosporella yangingensis CFCC 70738
Aplosporella yangingensis CFCC 70743
Aplosporella hesperidica CBS 208.37
Aplosporella hesperidica CBS 732.79
60/76/1 Aplosporella thailandica MFLU 16-0615
f Aplosporella artocarpi CPC 22791
aia Aplosporella ginkgonis CFCC 70746
Aplosporella ginkgonis CFCC 52442
Aplosporella ginkgonis CFCC 89661
Alanomyces indica CBS 134264
59/72/-
63/70/0.95
Outgroup
0.03
Figure 2. Phylogram generated from RAXML analysis based on ITS with tef1-a sequence data of Aplosporella isolates.
The tree was rooted in Alanomyces indica (CBS 134264). The MP, ML (2 50%), and BI (2 0.9) bootstrap supports are given
near the nodes, respectively. Isolates from this study are marked in blue, and ex-type strains are marked in bold.
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890 235
Yingying Wu et al.: New species and records of Botryosphaeriales
Dothiorella acericola CFCC 70755
99/1001 | Dothiorella acericola CFCC 70761
cove Dothiorella acericola KUMCC 18-0137
“ese Dothiorella acericola CFCC 70760
Dothiorella plurivora CBS 124724
80/97/Q.99f Dothiorella hortiarborum CFCC 70758
85/99/, a
Dothiorella hortiarborum CFCC 70759
Dothiorella hortiarborum CFCC 70756
56/33) ' Dothiorella hortiarborum CFCC 70757
55/5¢ Pp Dothiorella alpina CFCC 38299
LTL Dothiorella alpina CGMCC 3-18001
Dothiorella yannana CGMCC 3-17999
| Dothiorella magnoliae CFCC51563
Dothiorella yunnana CGMCC 3-18000
87/95/I Dothiorella rosulata CBS 121762
68/85/Q.92}
Dothiorella rosulata CBS 121761
ESE [I Dothiorella rosulata CBS 121760
Dothiorella rosulata CBS 500.72
La Tr Dothiorella mangifericola IRAN 1584C
752/0.97— Dothiorella mangifericola CBS 124727
79/861 Dothiorella citricola CBS 124729
anete Dothiorella citricola CBS 124728
be 96/100/1 fg9/95/9— Dothiorella westralis WA10NO01
5070/0 Dothiorella viticola CBS 117009
66/76/1
Dothiorella brevicollis CBS 130411
54/37/-
sv9s/I" Dothiorella striata CBS 124730
Dothiorella striata CBS 124731
Dothiorella heterophyllae CMW46458
=, Dothiorella saprophytica MFLUCC 23-0210
. Dothiorella longicollis CBS 122068
Dothiorella longicollis CBS 122067
Dothiorella longicollis CBS 122066
Dothiorella diospyricola CBS 145972
Dothiorella lampangensis MFLUCC 18-0232
Dothiorella obovata MFLUCC22-0058
Dothiorella tectonae MFLUCC18-0232
Dothiorella neclivorem DAR 80992
Pace 6p Dothiorella citrimurotticola CGMCC3.20392
abt aoe Dothiorella citrimurotticola CGMCC3.20394
Score Dothiorella uruguayensis CBS 124908
: Dothiorella oblonga CBS 121766
Dothiorella oblonga CBS 121765
15/05) LOMO. Dothiorella dulcispinae CBS 121764
100/100/1 Dothiorella dulcispinae CBS 130413
Dothiorella thailandica CBS 133991
Dothiorella albiziae MFLUCC 22-0057
' Dothiorella vinea-gemmae DAR 81012
Dothiorella omnivora CBS 392.80
Dothiorella omnivora CBS 124716
89/98/9,98) Dothiorella omnivora CBS 124717
, Dothiorella omnivora CBS 188.87
775/024" Dothiorella omnivora CBS 242.51
58/86/09] ful Dothiorella vidmadera CBS 725.79
Dothiorella vidmadera CBS 621.74
79/95/6911] ) Dothiorella parva CBS 124721
Dothiorella parva CBS 125580
Dothiorella guttulata MFLUCC 17-0242
67/67/2981 | Dothiorella parva CBS 124720
Dothiorella sarmentorum IMI 63581b
Dothiorella styphnolobii Cr01
Dothiorella americana CBS 128309
Dothiorella iberica CBS 115041
Dothiorella iberica CBS 113188
147, Dothiorella iberica CBS 113189
uae) L,I Dothiorella symphoricarpicola CPC 33923
79/77/09 ha Dothiorella californica CBS 119635
Dothiorella californica CBS 141587
ido) Dothiorella sempervirentis IRAN 1581C
Dothiorella sempervirentis IRAN 1583C
ae 3997/4 Dothiorella eriobotryae CBS140852
: 96/100/1 Dothiorella rhamni MFLUCC 14-0902
Dothiorella franceschinii CBS 147722
Dothiorella prunicola CBS 124723
Dothiorella santali WAC 13155
56/84. Dothiorella koae CMW 48017
Dothiorella moneti WAC 13154
TOI SS Dothiorella thripsita CBS 125445
100/100/1 Dothiorella pretoriensis CBS 130404
Dothiorella sp. CBS 121784
oo/1004| Dothiorella sp. CBS 121785
56/71/0.53 Dothiorella sp. CBS 121783
Dothiorella casuarinae CBS 120689
00/100 Dothiorella casuarinae CBS 120688
90/98/1 Dothiorella casuarinae CBS 120690
Dothiorella acacicola CBS 141295
187/008 100/100/1 Dothiorella capri-amissi CBS 121878
100/100/1 Dothiorella capri-amissi CBS 121763
100/100/1_ 4 Dothiorella baihuashanensis CFCC 58549
x2 Dothiorella baihuashanensis CFCC 58788
Dothiorella iranica CBS 124722
100/100/1 Dothiorella camelliae CMGCC 3.24158
Dothiorella zanthoxyli CMGCC 3.24159
Dothiorella ulmacea CBS 141414
100/100/1 Neofusicoccum parvum CMW9081
Neofusicoccum luteum CBS 562.92 Outgroup
0.03
Figure 3. Phylogram generated from RAXxML analysis based on ITS, tefl-a, and tub2 sequence data of Dothiorella iso-
lates. The tree was rooted in Neofusicoccum luteum (CBS 562.92) and Neofusicoccum parvum (CMW9081). The MP ML
(= 50%), and BI (= 0.9) bootstrap supports are given near the nodes, respectively. Isolates from this study are marked in
blue, and ex-type strains are marked in bold.
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890
Yingying Wu et al.: New species and records of Botryosphaeriales
Phaeobotryon negundinis CAA797
Phaeobotryon negundinis CAA798
Phaeobotryon negundinis CAA799
Phaeobotryon negundinis CPC 33388
96/95/1| Phaeobotryon negundinis CPC 33384
Phaeobotryon negundinis CPC 34752
Phaeobotryon negundinis MFLUCC 15-0436
95/99/1) Phaeobotryon juniperi JU 001
Phaeobotryon juniperi JU 005
i 0/100/11 Phaeobotryon cupressi CBS 124701
Phaeobotryon cupressi CBS 124700
94/100/1) Phaeobotryon platycladi CFCC 58800
Phaeobotryon platycladi CFCC58799
Phaeobotryon spiraeae CFCC 53926
87/98/0.97| Phaeobotryon spiraeae CFCC 53927
Phaeobotryon spiraeae CFCC 53925
99/100/1 Phaeobotryon mamane CPC 12442
Phaeobotryon mamane CPC 12440
Phaeobotryon rhoinum CFCC 52450
Phaeobotryon rhoinum CFCC 52449
98/100/1 Phaeobotryon aplosporum CFCC 53774
Phaeobotryon aplosporum CFCC 53776
86/99/1___| Phaeobotryon aplosporum CFCC 53775
Phaeobotryon aplosporum CFCC 58596
S3/520 BarranOe Phaeobotryon aplosporum CFCC58784
61/75/- | Phaeobotryon mali XJAU 2782
62/71/0.9N Phaeobotryon mali XJAU 2930
Phaeobotryon mali XJAU 2772
Phaeobotryon mali XJAU 3100
ena Phaeobotryon mali XJAU 3094
., Phaeobotryon rhois CFCC 89662
ae Phaeobotryon rhois CFCC 89663
100/100/1) Phaeobotryon fraxini CFCC 70762
Phaeobotryon fraxini CFCC 70763
Phaeobotryon ulmi CBS 123.30
Phaeobotryon ulmi CMH299
Phaeobotryon ulmi PB_11f
66/65/|' Phaeobotryon ulmi CBS 138854
99/99/1 Phaeobotryon ulmi CBS 174.63
Phaeobotryon ulmi CBS 114123
Phaeobotryon ulmi 94-13
Alanphillipsia aloeicola CBS 138896 Outgroup
66/62/0.96
0.02
Figure 4. Phylogram generated from RAxML analysis based on ITS, LSU, and tef1-a sequence data of Phaeobotryon iso-
lates. The tree was rooted in Alanphillipsia aloeicola (CBS 138896). The MP, ML (= 50%), and BI (= 0.9) bootstrap supports
are given near the nodes, respectively. Isolates from this study are marked in blue, and ex-type strains are marked in bold.
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890 997
Yingying Wu et al.: New species and records of Botryosphaeriales
Taxonomy
Aplosporella ginkgonis C.M. Tian, Z. Du & K.D. Hyde, Mycosphere 8(2): 1249
(2017)
Description. See Du et al. 2017.
Material examined. CHINA, Beijing City, Tongzhou District, Majugiao Wet-
land Park, 39°46'12"N, 116°37'12"E, on the disease branches of Cotinus
coggygria var. cinereus, 2 May 2023, Y.Y. Wu, BJUFC-S1931, living culture
CFCC 70746.
Notes. Aplosporella ginkgonis was first reported in Gansu Province, China,
causing canker and dieback disease in Ginkgo biloba and Morus alba (Du et
al. 2017). Zhu et al. (2018) and Li et al. (2023) discovered the species on Rhus
typhina and Zanthoxylum bungeanum, respectively, extending its host range.
In the present study, one isolate (CFCC 70746) was identified as A. ginkgonis
based on the phylogenetically highly supported clade with 99% MP, 95% ML,
and 0.94 BYPP values (Fig. 2) and morphological characteristics. This is the
first report of A. ginkgonis on Cotinus coggygria var. cinereus.
Aplosporella javeedii Jami, Gryzenh., Slippers & M.J. Wingf., Fungal Biology
118(2): 174 (2013)
Description. See Fan et al. 2015.
Material examined. CHINA, Beijing City, Tongzhou District, Hougezhuang
Plain Forest, 29°50'24"N, 116°54'00"E, on the dead branches of Styphnolo-
bium japonicum, 8 April 2023, C.M. Tian, S.J. Li & Y.Y. Wu, BUFC-S1932, liv-
ing culture CFCC 70733; ibid. on the dead branches of Forsythia suspensa,
BJFC-S1933, living culture CFCC 70734; ibid. on the dead branches of For-
sythia suspensa, BJFC-S1934, living culture CFCC 70735; ibid. on the dead
branches of Ulmus pumila, BJFC-S1935, living culture CFCC 70736; CHINA,
Beijing City, Tongzhou District, Central Green Forest Park, 39°52'16'N,
116°42'04"E, from branches of Acer truncatum, 12 April 2023, C.M. Tian,
Y.M. Liang, C. Peng, Y. Hu & Y.Y. Wu, BUFC-S1936, living culture CFCC 70737;
CHINA, Beijing City, Tongzhou District, Central Green Forest Park, 39°52'16'N,
116°42'04"E, on the dead branches of Sambucus williamsii, 19 April 2023,
C.M. Tian, C. Peng, R. Yuan, M.W. Zhang & Y.Y. Wu, BUFC-S1937, living culture
CFCC 70739; ibid. on the dead branches of Acer miyabei, BJFC-S1938, liv-
ing culture CFCC 70740; ibid. on the dead branches of Lagerstroemia indica,
BJFC-S19339, living culture CFCC 70741; ibid. on the dead branches of Xantho-
ceras sorbifolium, BJFC-S1940, living culture CFCC 70742; China, Beijing City,
Tongzhou District, Majugiao Wetland Park, 39°46'12"N, 116°37'12"E, from
branches of Syringa vulgaris, 2 May 2023, Y.Y. Wu, BUFC-S1941, living culture
CFCC 70744, ibid. on the dead branches of Ulmus pumila, BJFC-S1942, living
culture CFCC 70745.
Notes. Ap/osporella javeedii was initially reported on Celtis africana and Sear-
sia lancea in South Africa (Jami et al. 2014). Fan et al. (2015a) recorded this spe-
cies in China for the first time, associating it with the canker or dieback disease
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890 038
Yingying Wu et al.: New species and records of Botryosphaeriales
of five hosts: Albizia julibrissin, Broussonetia papyrifera, Gleditsia sinensis, Ju-
niperus chinensis, and Styphnolobium japonicum. Aplosporella javeedii is wide-
spread on host plants of more than 10 families (Fan et al. 2015a; Zhu et al.
2018; Pan et al. 2019; Lin et al. 2023a). In this study, we report new host records
for this species, including Acer miyabei, Acer truncatum, Forsythia suspensa,
Lagerstroemia indica, Sambucus williamsii, Syringa vulgaris, Ulmus pumila, and
Xanthoceras sorbifolium.
Aplosporella yangingensis L. Lin & X.L. Fan, MycoKeys 97: 9 (2023)
Description. See Lin et al. 2023a.
Material examined. CHINA, Beijing City, Tongzhou District, Central Green
Forest Park, 39°52'16"N, 116°42'04"E, on the dead branches of Acer trun-
catum, 12 April 2023, C.M. Tian, Y.M. Liang, C. Peng, Y. Hu & Y.Y. Wu,
BJFC-S1943, living culture CFCC 70743; ibid. BUFC-S1944, living culture
CFCC 70738.
Notes. Aplosporella yangingensis was first discovered on the branches of
Platycladus orientalis in Beijing (Lin et al. 2023a). In this study, the two iso-
lates (CFCC 70738 and CFCC 70743) from Acer truncatum formed a clade
with 100% MP, 100% ML, and 1.00 BYPP values in the multi-locus phyloge-
netic tree with A. yangingensis (Fig. 2). Compared with the description of Lin
et al. (2023a), this study has shorter conidia and thinner conidiogenous cells
(11.0-16.5 x 6.0-9.0 um vs. 16.0-21.5 x 6.0-9.5 um and 5.0-20.5 x 1.0-
2.0 um vs. 6.0-13.5 x 2.0-3.0 um). Thus, these isolates were identified as
A. yangingensis, and herewith we are providing a new host record for A. yan-
gingensis, Acer truncatum.
Dothiorella acericola Phookamsak, Tennakoon & K.D. Hyde, Fungal Diversity
95: 78 (2019)
Description. See Pan et al. 2021.
Material examined. CHINA, Beijing City, Tongzhou District, Hougezhuang
Plain Forest, 29°50'24"N, 116°54'00"E, on the dead branches of Forsythia sus-
pensa, 8 April 2023, C.M. Tian, S.J. Li & Y.Y. Wu, BUFC-S1948, living culture
CFCC 70755; CHINA, Beijing City, Tongzhou District, Majugiao Wetland Park,
39°46'12"N, 116°37'12"E, on the dead branches of Ginkgo biloba, 2 May 2023,
Y.Y. Wu, BUFC-S1949, living culture CFCC 70760; ibid. on the dead branches of
Syringa oblata, BJFC-S1950, living culture CFCC 70761.
Notes. Based on phylogenetic analyses (Fig. 3), three isolates in this study
clustered with Dothiorella acericola and formed a clade with 99% MP, 100% ML,
and 1.00 BYPP values. Dothiorella acericola is reported to be associated with
the canker disease of Acer palmatum in China (Phookamsak et al. 2019). Pan
et al. (2021, 2023) found that Do. acericola infests Ziziphus jujuba and Koelreu-
teria paniculata branches. The fungus was also recorded on dead branches of
Euonymus japonicus (Lin et al. 2023b). This is the first discovery of this fungus
in the host families Oleaceae and Ginkgoaceae.
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890 939
Yingying Wu et al.: New species and records of Botryosphaeriales
Dothiorella hortiarborum Y.Y. Wu & C.M. Tian, sp. nov.
MycoBank No: 851826
FIG=)
Etymology. “Hort” means “garden,” and “arbor” means “tree” in Latin. Collected
from Fraxinus chinensis and Lagerstroemia indica, both of which are landscap-
ing and greening trees.
Holotype. CHINA, Beijing City, Tongzhou District, Central Green Forest Park,
39°52'16"N, 116°42'04"E, on the dead branches of Fraxinus chinensis, 19 April
2023, C.M. Tian, C. Peng, R. Yuan, M.W. Zhang & Y.Y. Wu (holotype BUFC-S1951,
ex-type cultures CFCC 70756).
Description. Sexual morph: Not observed. Asexual morph: Conidiomata
pycnidial, scattered to aggregated, immersed to semi-immersed in bark, glo-
bose to subglobose, dark gray to black, unilocular, 260-450 um diam. Disc
black, ovoid, 310-330 um diam. Ostioles single, light gray, circular, central,
papillate, 30-45 um diam. Locules single, black, oval, 100-380 um, Conidio-
phores reduced to conidiogenous cells. Conidiogenous cells: hyaline, smooth,
thin-walled, holoblastic, cylindrical to subcylindrical, 4.5-11.0 x 2.0-4.0 um (av.
+ S.D.= 6.8 + 1.3 x 2.9 + 0.5 um). Conidia initially hyaline, then producing light
yellow pigmentation, uneven surface, thick-walled, dark brown when matrues,
1-septate, constricted at the septum, smooth, ovoid with a broadly rounded apex,
truncate base. 10.0-19.0 x 6.0-11.0 um (av. + S.D.= 14.9 + 2.6 x 8.1 + 1.0 um).
Figure 5. Dothiorella hortiarborum (BJFC-S1951) A, B habit of conidiomata on branch C transverse section of conidioma
D longitudinal section through conidioma E, F conidiogenous cells and conidia G top (left) and bottom (right) sides of
colonies on potato dextrose agar (PDA) H, I conidia. Scale bars: 1000 ym (A); 200 um (B-D); 10 um (E-F, H-1).
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890 240
Yingying Wu et al.: New species and records of Botryosphaeriales
Culture characters. Colonies on PDA with aerial mycelium gray-green, thick
and dense, fluffly, margin with undulate and irregular, reverse with inky blue pig-
ment accumulation, reaching 60 mm diam in 7 days at 25 °C.
Other material examined. CHINA, Beijing City, Tongzhou District, Central Green
Forest Park, 39°52'16'N, 116°42'04"E, on the dead branches of Fraxinus chinensis,
19 April 2023, C.M. Tian, C. Peng, R. Yuan, M.W. Zhang & Y.Y. Wu, BUFC-S2366, liv-
ing culture CFCC 70757; CHINA, Beijing City, Tongzhou District, Central Green For-
est Park, 39°52'16'"N, 116°42'04'E, on the dead branches of Lagerstroemia indica,
19 April 2023, C.M. Tian, C. Peng, R. Yuan, M.W. Zhang & Y.Y. Wu, BUFC-S1952,
living culture CFCC 70758; ibid. BJFC-S236/7, living culture CFCC 70759.
Notes. Dothiorella hortiarborum formed an independent clade with 87% MP
97% ML, and 0.99 BYPP values and is distinct from Do. acericola and Do. pluriv-
ora in the multi-locus analyses (Fig. 3). Morphologically, Do. hortiarborum can be
distinguished from Do. acericola by shorter conidia (Phookamsak et al. 2019) and
Do. plurivora by smaller conidia (10.0-19.0 x 6.0-11.0 um vs. 22.3-22.7 x 10.8-
11.2 um) (Abdollahzadeh et al. 2014). Additionally, Do. hortiarborum differs from
Do. acericola in tef1-a (five bp difference from 170 characters, with 97.1% simi-
larity, including no gaps) sequences, and Do. plurivora in tef1-a (one bp difference
from 254 characters, with 99.6% similarity, including one gap), tub2 (three bp dif-
ference from 370 characters, with 99.2% similarity, including one gap) sequences.
Phaeobotryon fraxini Y.Y. Wu & C.M. Tian, sp. nov.
MycoBank No: 851827
Fig. 6
Etymology. Named after the host, Fraxinus chinensis.
Holotype. CHINA, Beijing City, Tongzhou District, Central Green Forest Park,
39°52'16"N, 116°42'04"E, on the dead branches of Fraxinus chinensis, 19 April
2023, C.M. Tian, C. Peng, R. Yuan, M.W. Zhang & Y.Y. Wu (holotype BUFC-S1953,
ex-type cultures CFCC 70762).
Description. Sexual morph: Not observed. Asexual morph: Conidiomata pycnid-
ial, scattered, occasionally aggregated, superficial or immersed, globose, dark
brown to black, unilocular, 200-360 um diam. Disc inconspicuous. Ostioles single,
brown or black, circular, central, papillate, 40-85 um diam. Locules single, globose,
100-170 um, Conidiophores reduced to conidiogenous cells. Conidiogenous cells
hyaline, smooth, thin-walled, holoblastic, cylindrical, formed from the cells lining
the inner walls of the locules, 7.0-14.0 x 1.0-5.0 um (av. + S.D.= 10.6 + 2.0 x 3.1
+ 0.8 um). Conidia initially hyaline, smooth, thin-walled, then gradually producing
light yellow pigment, becoming yellow or light brown, occasionally with bubbles,
mature with 1-septate, brownish yellow to dark brown, oblong, obtuse, rounded at
both ends, 13.0-20.0 x 7.0-10.0 um (av. + S.D.= 17.6 + 1.3 x 8.7 + 0.7 um).
Culture characters. Colonies on PDA with aerial gray-white mycelium, thick and
dark black at the edge, thin and paler in color in the center, fluffly, entire margin, re-
verse with black pigment accumulation, reaching 60 mm diam in 7 days at 25 °C.
Other material examined. CHINA, Beijing City, Tongzhou District, Central
Green Forest Park, 39°52'16"N, 116°42'04"E, on the dead branches of Fraxinus
chinensis, 19 April 2023, C.M. Tian, C. Peng, R. Yuan, M.W. Zhang & Y.Y. Wu,
BJFC-S2368, living culture CFCC 70763.
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890 oA
Yingying Wu et al.: New species and records of Botryosphaeriales
C longitudinal section through conidioma D, E conidiogenous cells and conidia F top (left) and bottom (right) sides of
colonies on potato dextrose agar (PDA) G-L conidia. Scale bars: 500 um (A); 200 um (B, C); 10 um (D, E, G-L).
Notes. Based on multi-locus phylogenetic analysis, the two isolates cluster
separately in a high-supported clade with 100% MP, 100% ML, and 1.00 BYPP
value (Fig. 4). In the phylogenetic analysis, Phaeobotryon fraxini showed a close
relationship to P mali and P. rhois. These three species could be distinguished
based on ITS, tef1-a, and LSU loci from P. mali by nineteen bp (6/465 in ITS;
10/184 in tef1-a; 3/559 in LSU) and P. rhois by twenty-two bp (7/465 in ITS;
12/184 in tef1-a; 3/559 in LSU). Moreover, P. fraxini differs from P mali and
P. rhois in having smaller conidia (13.0-20.0 x 7.0-10.0 um vs. 22.0-31.5 x
12-16.5 um for P mali and 20-25 x 10-12 um for P rhois) (Fan et al. 2015b;
Jia et al. 2023) (Table 3). Therefore, P. fraxini is introduced as a novel species.
Table 3. Comparison of species in Phaeobotryon.
Species
Phaeobotryon aplosporum
P mali
P cupressi
P. fraxini
P juniperi
P mamane
P. negundinis
P platycladi
P rhoinum
P rhois
P spiraeae
P ulmi
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890
Host
Rhus typhina
Malus pumila
Cupressus sempervirens
Fraxinus chinensis
Juniperus formosana
Sophora chrysophylla
Acer negundo
Platycladus orientalis
Rhus typhina
Rhus typhina
Spiraea salicifolia
Ulmus laevis
Location
China
China
Iran
China
China
USA
Russia
China
China
China
China
Germany
Conidial size
17-19 x 5.5-7
22.0-31U5 <4 2516.5
24.1525 *12.2-12:5
13-20 x 7-10
245 = 27.5% 412.0—13:5
35-38 x 14-15
16-24.5 x 7.9-11.5
23.0-31.0 x 9.5-12.5
19-21 x 7.5-9
20-25 x 10-12
2315-28 585-135
28.5-32.5 x 16.5-18.5
Septation
aseptate
1-septate
1(-2)-septate
1-septate
1-septate
1(-2)-septate
aseptate
aseptate or 1-septate
1-septate
1-septate
aseptate
aseptate or 1-septate
Reference
Pan et al. 2019
Jia et al. 2023
Abdollahzadeh et al. 2009
This study
Peng et al. 2023
Phillips et al. 2008
Daranagama et al. 2016
Lin et al. 2023a
Zhu et al. 2018
Fan et al. 2015b
Jin and Karunarathna 2021
Zhang et al. 2021
242
Yingying Wu et al.: New species and records of Botryosphaeriales
Discussion
In this paper, 23 Botryosphaeriales isolates were identified as six species
based on multi-locus phylogenetic analyses. These species included two new
species, namely Dothiorella hortiarborum and Phaeobotryon fraxini, and four
new hosts: Aplosporella ginkgonis on Cotinus coggygria var. cinereus; A. javee-
dii on Acer miyabei; Acer truncatum; Forsythia suspensa; Lagerstroemia indica;
Sambucus williamsii, Syringa vulgaris; Ulmus pumila; Xanthoceras sorbifolium;
A. yangingensis on Acer truncatum; and Do. acericola on Forsythia suspensa;
Ginkgo biloba; and Syringa oblata. The six fungal species identified in this study
involve a total of 13 different hosts, which elucidates the wide range of hosts
of Botryospaeriales.
Aplosporella is the type genus of Aplosporellaceae (Slippers et al. 2013).
The distinctive morphological feature of Aplosporella species is that both as-
cospores and conidia are aseptately hyaline to pigmented (Slippers et al. 2013;
Phillips et al. 2019). In this study, a total of three new host record species of the
genus were identified, including A. ginkgonis, A. javeedii, and A. yangingensis.
Aplosporella javeedii has the highest isolation rate and the widest host range,
involving five orders of host plants, including Dipsacales, Fabales, Lamiales,
Myrtales, and Rosales. Currently, this species is mainly found in warm temper-
ate and tropical regions (Fan et al. 2015a; Zhu et al. 2018), and further explo-
ration is needed to determine whether the geographic range of A. javeedii is
related to climate.
Dothiorella was considered a synonym of Diplodia based on a broad mor-
phological concept (Crous and Palm 1999). Phillips et al. (2005) compared the
morphological characteristics again and found that the conidia of Dothiorella
were brown, with 1-septate in early development, and the conidia still adhered
to the conidiogenous cells. In contrast, the conidia of Diplodia become black
and septate after being excreted from the conidiomata. Crous et al. (2006) con-
firmed these morphological differences. Therefore, Dothiorella is regarded as
an independent genus in the Botryosphaeriaceae. In this study, the conidia of
Do. hortiarborum are transparent and aseptate when attached to conidioge-
nous cells. After being released by the conidiomata, the conidia bear yellowish
pigment or become brown with a 1-septate. In recent years, many new species
of Dothiorella have been published with conidial morphology similar to Do. hor-
tiarborum (Li et al. 2023; Lin et al. 2023a; Wu et al. 2023). These suggest that
the morphological characteristics of Dothiorella are not always stable. Thus,
it is not accurate to rely solely on the morphology of conidia for Dothiorella;
combining phylogenetic analysis and the size of conidia of neighboring spe-
cies is necessary. Dothiorella species have been reported on more than 20 host
plants in China (https://fungi.ars.usda.gov/). This study has expanded its host
range in Oleaceae plants (Do. acericola in Forsythia suspensa, Ginkgo biloba
and S. oblata, and Do. hortiarborum in Fraxinus chinensis).
Currently, many Dothiorella species have been recorded from Fraxinus, distrib-
uted mainly in regions such as Europe and North America (Table 4). In this study, a
new species, Do. hortiarborum, from F. chinensis, was introduced in China. Howev-
er, based on morphological and DNA sequence data, Do. hortiarborum shows sig-
nificant differences from other species in Fraxinus. Phylogenetic analysis showed
that Do. hortiarborum belongs to a different lineage from Do. omnivora, Do. sp.,
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890 943
Yingying Wu et al.: New species and records of Botryosphaeriales
Table 4. Comparison of species from Fraxinus in Dothiorella.
Specise Host Location Conidial size Septation Reference
Dothiorella concaviuscula Fraxinus viridis USA 4-6 x 2.5-3 no description Jepson 1896
Do. fraxini Fraxinus sp. Belgium 26-30 x 12 1-septate Saccardo 1892
Do. fraxinicola Fraxinus sp. USA 18-30 x 6-7 no description Ellis and Everhart 1895
Do. hortiarborum Fraxinus chinensis China 10.0-19.0 x 6.0-11.0 1-septate This study
Do. omnivora Fraxinus excelsior Bosnia 19.3-25.5 x 7.5-10.6 1-septate Linaldeddu et al. 2016
Do. sp. Fraxinus excelsior Bosnia, Herzegovina 11-14x 6-8 2-4-septate Zlatkovié et al. 2016
Do. vidmadera Fraxinus ornus Australia 21.2-21.9 x 9.6-9.8 1-septate Pitt et al. 2013
and Do. vidmadera (Fig. 3), while distinguishing them based on the size of conidia
and the number of septates (Table 4). Do. concaviuscula, Do. fraxini, and Do. frax-
inicola were not available for sequence information due to their earlier publication;
however, Do. hortiarborum can also be easily distinguished from them based on
their documented conidia size. In addition, Do. lagerstroemiae and Do. hortiarbo-
rum were both isolated from Lagersiroemia alba, but its conidia were significantly
smaller than Do. hortiarborum (8.3-10 x 3.5—4 um vs. 10.0-19.0 x 6.0-11.0 um).
Phaeobotryon species have more overlapping morphological characters,
with 1(—2) septate or aseptate conidia and similar pigmentation variations. For
example, P cupressi and P juniperi have overlapping sizes of conidia (24.1-
25 x 12.2-12.5 um vs. 24.5-27.5 x 12.0-13.5 pm), P rhoinum and P rhois are
derived from the same host and geographic origin, and the conidia have 1-sep-
tate (Table 3). So, morphology combined with phylogenetics to further clarify
the affinities between species is essential. Furthermore, Phaeobotryon species
were reported on a variety of hosts and considered to be potential or oppor-
tunistic pathogens (Weiland et al. 2016; Zhu et al. 2020; Ilyukhin and Ellouze
2023; Jia et al. 2023). In this study, P fraxini was isolated only from dead Frax-
inus chinensis; more extensive specimen collection was needed to confirm its
distribution characteristics and pathogenicity.
Although Botryosphaeriales recorded many fungi on Index Fungorum
(https://www.indexfungorum.org/), only some species are now recognized.
Mainly due to the early records of many species, the lack of model specimens,
or the low quality of specimens, it is difficult to obtain strains and DNA data.
Therefore, more detailed sampling is needed to revise the classification system
of related taxa in Botryosphaeriales.
Additional information
Conflict of interest
The authors have declared that no competing interests exist.
Ethical statement
No ethical statement was reported.
Funding
This study is financed by National Natural Science Foundation of China (Project No.:
32371887), Survey of Insect and Pathogen Diversity in Beijing Municipal Administra-
tive Center.
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890 oA
Yingying Wu et al.: New species and records of Botryosphaeriales
Author contributions
Conceptualization, Yingying Wu and Chengming Tian; data curation, Yingying Wu; funding
acquisition, Chengming Tian; investigation, Yingying Wu, Cheng Peng, Rong Yuan, Mingwei
Zhang, Yang Hu; project administration, Chengming Tian; resources, Yingying Wu, Cheng
Peng, Rong Yuan, Mingwei Zhang, Yang Hu; supervision, Chengming Tian; writing-original
draft, Yingying Wu; writing-review and editing, Yingying Wu, Cheng Peng, and Chengming
Tian. All authors have read and agreed to the published version of the manuscript.
Author ORCIDs
Yingying Wu © https://orcid.org/0009-0007-5095-2738
Rong Yuan ® https://orcid.org/0009-0006-5597-7531
Chengming Tian © https://orcid.org/0000-0002-3352-7664
Data availability
All of the data that support the findings of this study are available in the main text or
Supplementary Information.
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Supplementary material 1
Aplosporella
Authors: Yingying Wu, Cheng Peng, Rong Yuan, Mingwei Zhang, Yang Hu, Chengming Tian
Data type: pdf
Copyright notice: This dataset is made available under the Open Database License
(http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License
(ODbL) is a license agreement intended to allow users to freely share, modify, and
use this Dataset while maintaining this same freedom for others, provided that the
original source and author(s) are credited.
Link: https://doi.org/10.3897/mycokeys.106.122890.suppl1
Supplementary material 2
Dothiorella
Authors: Yingying Wu, Cheng Peng, Rong Yuan, Mingwei Zhang, Yang Hu, Chengming Tian
Data type: pdf
Copyright notice: This dataset is made available under the Open Database License
(http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License
(ODbL) is a license agreement intended to allow users to freely share, modify, and
use this Dataset while maintaining this same freedom for others, provided that the
original source and author(s) are credited.
Link: https://doi.org/10.3897/mycokeys.106.122890.suppl2
Supplementary material 3
Phaeobotryon
Authors: Yingying Wu, Cheng Peng, Rong Yuan, Mingwei Zhang, Yang Hu, Chengming Tian
Data type: pdf
Copyright notice: This dataset is made available under the Open Database License
(http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License
(ODbL) is a license agreement intended to allow users to freely share, modify, and
use this Dataset while maintaining this same freedom for others, provided that the
original source and author(s) are credited.
Link: https://doi.org/10.3897/mycokeys.106.122890.suppl3
MycoKeys 106: 225-250 (2024), DOI: 10.3897/mycokeys.106.122890 250
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